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首页> 外文期刊>Journal of Clinical Microbiology >Immunofluorescence assay for human immunodeficiency virus antibody: investigation of cell fixation for virus inactivation and antigen preservation.
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Immunofluorescence assay for human immunodeficiency virus antibody: investigation of cell fixation for virus inactivation and antigen preservation.

机译：人类免疫缺陷病毒抗体的免疫荧光测定：用于病毒灭活和抗原保存的细胞固定研究。

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Four cell fixation procedures were investigated for their abilities to inactivate human immunodeficiency virus (HIV) and preserve its antigenicity for antibody detection by immunofluorescence in MOLT-4-T4 cells. Air-dried cell smears were fixed in cold acetone, in acetone-methanol (1:1), in acetone-methanol (1:1) followed by 70% ethanol and then methanol, or in paraformaldehyde-acetone. Acetone alone did not inactivate cell-associated HIV, but the other three procedures did. HIV inactivation was achieved by storage of acetone-fixed cells at -70 degrees for 40 days. Antigenicity was measured by immunofluorescence assay titrations of selected human sera, a cerebrospinal fluid, and a gp41 monoclonal antibody. Acetone provided the best fixation as measured by fluorescence intensity and antibody titers. The other fixation methods all yielded weaker fluorescence signals and/or decreased titers. Acetone fixation and storage for 40 days at -70 degrees C provides safe and accurate immunofluorescence assay reagents.
机译：研究了四种细胞固定程序灭活人类免疫缺陷病毒（HIV）并保留其抗原性以通过MOLT-4-T4细胞中的免疫荧光进行抗体检测的能力。风干的细胞涂片固定在冷丙酮，丙酮-甲醇（1：1），丙酮-甲醇（1：1），70％乙醇，然后是甲醇或多聚甲醛-丙酮中。单独使用丙酮并不能使与细胞相关的HIV灭活，但其他三种方法却可以。通过将丙酮固定的细胞在-70度下存储40天来实现HIV灭活。通过对选定的人血清，脑脊液和gp41单克隆抗体进行免疫荧光测定滴定来测量抗原性。通过荧光强度和抗体效价测定，丙酮提供了最佳的固定。其他固定方法均产生较弱的荧光信号和/或降低的滴度。丙酮在-70摄氏度下固定并保存40天，可提供安全，准确的免疫荧光测定试剂。

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